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There has recently been an explosion of studies implicating the human microbiome in playing a critical role in many disease and wellness states. The etiology of abnormal semen analysis (SA) parameters is not identified in 30% of cases; investigations involving the semen microbiome may bridge this gap. Here, we explore the relationship between the semen microbiome and alterations of sperm parameters. We recruited men presenting for fertility evaluation or vasectomy consultation with proven biological paternity. SA and next generation sequencing was performed. Differential abundance testing using Analysis of composition of Microbiota with Bias Correction (ANCOM-BC) was performed along with canonical correlational analysis for microbial community profiling. Men with abnormal (N = 27) sperm motility showed a higher abundance of Lactobacillus iners compared to those with normal (N = 46) sperm motility (mean proportion 9.4% versus 2.6%, p = 0.046). This relationship persisted on canonical correlational analysis (r = 0.392, p = 0.011). Men with abnormal sperm concentration (N = 20) showed a higher abundance of Pseudomonas stutzeri (2.1% versus 1.0%, p = 0.024) and Pseudomonas fluorescens (0.9% versus 0.7%, p = 0.010), but a lower abundance of Pseudomonas putida (0.5% versus 0.8%, p = 0.020), compared to those with normal sperm concentration (N = 53). Major limitations are related to study design (cross-sectional, observational). Our results suggest that a small group of microorganisms may play a critical role in observed perturbations of SA parameters. Some of these microbes, most notably Lactobacillus iners, have been described extensively within other, fertility-related, contexts, whereas for others, this is the first report where they have potentially been implicated. Advances in our understanding of the semen microbiome may contribute to potentially new therapeutic avenues for correcting impairments in sperm parameters and improving male fertility.

Sperm capture techniques that immobilize sperm to halt their motility are essential for the long-term analysis of individual sperm. These techniques are beneficial in assisted reproductive technologies such as intracytoplasmic sperm injection (ICSI) by allowing selective screening of sperm. However, there is a notable lack of high-throughput and non-destructive sperm capture methods that allow the flagellum to beat freely, which is crucial for accurately reflecting the behavior of unfettered, freely swimming sperm. To bridge this gap, we introduce a novel microfluidic device specifically engineered to capture sperm without restricting flagellar motion. The design utilizes sperm's innate boundary-following behavior in both 3D and 2D environments to direct them into a capture zone. Once captured, the sperm head is restrained while the flagellum remains free to exhibit natural beating patterns. Utilizing this device, we explore the effects of hyperactivating agents, temperature, and their combined influence on the dynamics of bovine sperm flagella. The unrestricted flagellar motion offered by our device yields two prominent advantages: it mirrors the flagellar behavior of free-swimming sperm, ensuring research findings are consistent with natural sperm activity, and it prevents imaging overlap between the flagellum and the capture structures, simplifying the automation of flagellar tracking and analysis. This technological advancement facilitates the collection of waveform parameters along the entire flagellum, addressing inconsistencies that have arisen in previous research due to differing measurement sites, and enabling precise extraction of sperm behavioral properties. [ABSTRACT FROM AUTHOR]

As the global male infertility rate continues to rise, there is an urgent imperative to investigate the underlying causes of sustained deterioration in sperm quality. The gut microbiota emerges as a pivotal factor in host health regulation, with mounting evidence highlighting its dual influence on semen. This review underscores the interplay between the Testis-Gut microbiota axis and its consequential effects on sperm. Potential mechanisms driving the dual impact of gut microbiota on sperm encompass immune modulation, inflammatory responses mediated by endotoxins, oxidative stress, antioxidant defenses, gut microbiota-derived metabolites, epigenetic modifications, regulatory sex hormone signaling. Interventions such as probiotics, prebiotics, synbiotics, fecal microbiota transplantation, and Traditional natural herbal extracts are hypothesized to rectify dysbiosis, offering avenues to modulate gut microbiota and enhance Spermatogenesis and motility. Future investigations should delve into elucidating the mechanisms and foundational principles governing the interaction between gut microbiota and sperm within the Testis-Gut microbiota Axis. Understanding and modulating the Testis-Gut microbiota Axis may yield novel therapeutic strategies to enhance male fertility and combat the global decline in sperm quality. [ABSTRACT FROM AUTHOR]

Heat stress (HS) has become a common stressor, owing to the increasing frequency of extreme high-temperature weather triggered by global warming, which has seriously affected the reproductive capacity of important livestock such as sheep. However, little is known about whether HS reduces sperm motility by inducing circadian rhythm disorders in rumen microorganisms and metabolites in sheep. In this study, the year-round reproduction of two-year-old Hu rams was selected, and the samples were collected in May and July 2022 at average environmental temperatures between 18.71 °C and 33.58 °C, respectively. The experiment revealed that the mean temperature-humidity index was 86.34 in July, indicating that Hu rams suffered from HS. Our research revealed that HS significantly decreased sperm motility in Hu rams. Microbiome analysis further revealed that HS reshaped the composition and circadian rhythm of rumen microorganisms, leading to the circadian disruption of microorganisms that drive cortisol and testosterone synthesis. Serum indicators further confirmed that HS significantly increased the concentrations of cortisol during the daytime and decreased the testosterone concentration at the highest body temperature. Untargeted metabolomics analysis revealed that the circadian rhythm of rumen fluid metabolites in the HS group was enriched by the cortisol and steroid synthesis pathways. Moreover, HS downregulated metabolites, such as kaempferol and L-tryptophan in rumen fluid and seminal plasma, which are associated with promotion of spermatogenesis and sperm motility; furthermore, these metabolites were found to be strongly positively correlated with Veillonellaceae_UCG_001. Overall, this study revealed the relationship between the HS-induced circadian rhythm disruption of rumen microorganisms and metabolites and sperm motility decline. Our findings provide a new perspective for further interventions in enhancing sheep sperm motility with regard to the circadian time scale. [ABSTRACT FROM AUTHOR]

Purpose: The impact of air pollution on semen quality has been confirmed, yet the joint effect remains unclear. We evaluate the individual and joint associations of particulate (PM2.5 and PM10) and gaseous pollutants (NO2, SO2, O3 and CO) with semen quality. Methods: We included 5,114 men in this study from 2014 to 2022. The individual and joint associations were measured by multiple linear regression models. Results: Sperm motility and semen volume were inversely associated with pollutant concentrations during every stage of sperm development, especially at lag days 0–9 and 10–14 (all P < 0.05). Stratified analyses showed that the study pollutants (except CO) had a positive effect on semen concentration during the stage of sperm development, especially in spring and autumn, while a decreased total sperm number was associated with CO (all P < 0.05). However, joint associations of particulate and gaseous pollutants with semen quality parameters were not statistically significant (all P > 0.05). Conclusions: During all stages of sperm development, particulate and gaseous pollutants had individual negative impacts on sperm motility and semen volume, and these impacts were less pronounced in spring and autumn. Our findings highlight the importance and necessity of reducing the exposure to pollutants especially in the critical stage of sperm development to improve semen quality. [ABSTRACT FROM AUTHOR]

Simple Summary: Artificial insemination (AI) is crucial for dairy cattle worldwide due to its impact on reproductive efficiency and profitability. This study examined the effects of thawing time and temperature, combined with a thermo-resistance test (TRT), on sperm motility and kinematic variables in dairy bulls' semen for AI. The semen thawing process is critical in maintaining sperm quality, requiring strict adherence to standardized techniques. Incorrect thawing procedures can damage sperm structures. Rapid thawing is necessary to prevent ice recrystallization, and excessive heat exposure can lead to detrimental changes. The thermo-resistance test simulates conditions of the female reproductive tract to assess sperm longevity. Results indicated that Jersey bulls were greater for total and progressive motility percentages compared to Holstein bulls. Jersey bulls exhibited higher values for various velocity parameters and linearity, while Holstein bulls had a lower crossover frequency. The optimal thawing condition was 37 °C for 30 s. Sperm quality decreased with longer post-thawing times. This investigation aimed to analyze the effect that thawing time and temperature in combination with a termo-resistance test had on straws from dairy bulls used for artificial insemination (AI) on semen motility and kinematic variables measured with CASA systems. Eight animals of Holstein and Jersey breeds were used, and nine frozen-thawed semen doses per animal were analyzed for each breed. Three temperatures (35, 37, and 40 °C) and three thawing times (35, 40, and 45 s) were evaluated using a factorial design. Motility and kinematic patterns were analyzed using CASA-mot (Computer-Assisted Semen Analysis of motility) technology at different post-thawing times (0.5, 1, and 2 h). Sperm motility in Jersey bulls was higher (p < 0.05) than in Holstein ones (64.52 ± 1.45% and 53.10 ± 1.40%, respectively). The same effect was seen with progressive motility among the two breeds (Jersey: 45.29 ± 1.00%; Holstein: 36.30 ± 0.98%, p < 0.05). The Jersey breed presented higher values (p < 0.05) of curvilinear velocity (VCL), rectilinear velocity (VSL), average velocity (VAP), linearity on forward progression (LIN), and wobble (WOB). The Holstein breed showed a lower mean value (p < 0.05) of the beat-cross frequency (BCF) compared to the Jersey breed, thus suggesting an effect on VCL and VAP. During the post-thaw period, a gradual increase in VCL was observed at 2 h. VSL and VAP showed a decrease (p < 0.05) as the post-thaw period was prolonged. The study showed differences in sperm quality between Holstein and Jersey breeds, influenced by cryopreservation, thawing, and post-thawing incubation. Thawing at 37 °C for 30 s was considered optimal in relation to sperm motility. In addition, a decrease in sperm quality was observed as post-thawing time increased. [ABSTRACT FROM AUTHOR]

Semen preservation quality affects the artificial insemination success rate, and seminal exosomes are rich in various proteins that are transferable to sperm and conducive to sperm-function preservation during storage. However, the specific effects of these proteins remain unclear. In this study, the specific effects of these proteins on semen preservation quality and fertilization capacity were investigated through a proteomic analysis of seminal exosomes from boars with high conception rates (HCRs) and low conception rates (LCRs). The results revealed significant differences in the expression of 161 proteins between the two groups, with the GPX5 level being significantly higher in the HCR group (p < 0.05). The role of GPX5 was further investigated by constructing engineered exosomes enriched with GPX5 (Exo-GPX5), which could successfully transfer GPX5 to sperm. Compared to the control group, Exo-GPX5 could significantly improve sperm motility on storage days 4 and 5 and enhance the acrosome integrity on day 5 (p < 0.05). Additionally, Exo-GPX5 increased the total antioxidant capacity (T-AOC) of sperm, reduced the malondialdehyde (MDA) level, and decreased the expression of antioxidant proteins SOD1 and CAT (p < 0.05). In simulated fertilization experiments, Exo-GPX5-treated sperm exhibited higher capacitation ability and a significant increase in the acrosome reaction rate (p < 0.05). Overall, Exo-GPX5 can improve boar semen quality under 17 °C storage conditions and enhance sperm fertilization capacity. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the effect of silver nanoparticles (AgNPs) on donkey sperm parameters and ultrastructure. AgNPs were synthesized, purified and resuspended in the extender. Nine frozen–thawed donkey sperm samples were exposed to different concentrations of AgNPs (0, 1.25, 2.5, 5, 12.5, 25 and 50 μg/mL). Sperm parameters: total (TMOT, %) and progressive (PMOT, %) sperm motility, plasma (LIVE, %) and acrosomal membrane integrity (AIS, %), and sperm morphology (MORF, %) were evaluated immediately after AgNPs exposure (T0) and after 2 h of incubation (T2). The interaction beween AgNPs and spermatozoa was visualized by transmission electron microscopy (TEM). At T0, sperm motility and AIS were reduced (p <.05) when using concentrations ≥50 and ≥25 μg/mL, respectively. At T2, sperm motility and LIVE were significantly decreased (p <.05) in concentrations ≥25 and ≥50 μg/mL, respectively. TEM analysis revealed nanoparticle adhesion to the acrosomal region of the plasma membrane. In conclusion, AgNPs at concentrations ≥25 μg/mL impair motility, acrosome and plasma membrane integrity of donkey sperm, which may be mediated by adhesion to the acrosomal region of the sperm surface membrane. [ABSTRACT FROM AUTHOR]

The aim of this experiment was to assess the effect of media viscosity on ram sperm motility, kinematics and rheotaxis in vitro by using methylcellulose as a media thickener. Frozen–thawed semen of three rams was thawed and diluted in Tyrode's albumin lactate pyruvate (TALP) media supplemented with 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% and 0.7% w/v of methylcellulose. Sperm motility and kinematic characteristics were analysed using computer‐assisted sperm analysis (CASA). The rheotactic behaviour was assessed in a microfluidic channel, and the number of spermatozoa that passed the 10 mm point of a microfluidic channel over a 2min period against a flow rate of 30 μm/sec was assessed. The use of media with higher viscosity (higher levels of methylcellulose) resulted in significantly lower (p <.05) sperm motility and kinematic parameters. Moreover, higher levels of methylcellulose reduced (p <.05) the number of spermatozoa that exhibited positive rheotaxis. In conclusion, viscosity affected the kinematic properties and rheotactic behaviour of ram sperm. [ABSTRACT FROM AUTHOR]

β‐nerve growth factor (βNGF) plays a crucial role in reproductive physiology and sperm quality. Enzymatic activity of seminal plasma and vaginal fluids reduces available βNGF and it has been demonstrated that chitosan microspheres could protect rrβNGF from degradation. This study examined the effects of microencapsulated rrbNGF with chitosan on rabbit sperm viability, motility and capacitation status. Results showed that 0.5 and 1 μg/mL of microencapsulated rrβNGF, as well as free rrβNGF or empty microspheres, did not adversely affect sperm viability or total motility after 2 h of incubation. However, the highest progressivity kinetic parameters were observed with 1 μg/mL free rrβNGF, while the highest curvilinear velocity (VCL) occurred with 0.5 μg/mL microencapsulated rrβNGF. Empty chitosan microspheres did not induce acrosome reaction (AR), but both concentrations of free and rrβNGFch favoured AR during in vitro incubation. The study suggests that using chitosan spheres did not show any adverse effects on sperm traits, unlike free rβNGF and rrβNGFch promoted capacitation and AR. Further research is needed to explore the potential of rrβNGFch in modifying in vitro capacitation and inducing ovulation during artificial insemination. [ABSTRACT FROM AUTHOR]

A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT‐TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen–thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT‐TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT‐TALP or INRA96 with PHE at 30 or 38.2°C for 22 h. Following incubation, total motility (TM), progressive motility (PM), viability and acrosome integrity were evaluated. The results showed that TM was significantly higher (p <.001) at 30°C in both media, while PM was higher for INRA96 at 30°C compared to 38°C (p <.05). Moreover, INRA96 at 30°C exhibited higher sperm viability and acrosome integrity (p <.001) compared to the other experimental groups. These preliminary results suggest that incubating thawed equine spermatozoa at 30°C with PHE in INRA96 successfully maintains motility, viability and acrosome integrity in equine spermatozoa, indicating its potential use for conventional equine IVF. [ABSTRACT FROM AUTHOR]

The aim was to evaluate the influence of residual feed intake (RFI) on the semen characteristics of Dorper sheep after cryopreservation. Six Dorper sheep were used, classified from a population of 64 according to feed efficiency (groups of animals-positive RFI and negative RFI). The ejaculates were harvested three times a week, with a minimum interval of 48-h between collections, being seven collections per animal, totalling 42 ejaculates. Motility, plasma membrane integrity, acrosomal, mitochondrial activity and perivitelline membrane binding tests were performed. Negative RFI animals presented higher progressive motility (31.28%) than of RFI positive (25.37%). Negative and positive RFI animals present similar qualities in the assessment of plasma and acrosomal membrane integrity and mitochondrial activity, and negative RFI animals presented a higher number of membrane-bound spermatozoa than the positive RFI animals. Negative RFI animals displayed better rates of sperm motility after thawing, and are more efficient in the fertilisation in vitro. RFI did not influence the integrity of the plasma and acrosomal membrane, nor the mitochondrial activity of Dorper sheep spermatozoa after cryopreservation. [ABSTRACT FROM AUTHOR]

Cisplatin (a platinum-based drug) has chemotherapeutical potency in treating different kinds of cancer but elicits side effects of testicular dysfunction while hesperetin has antioxidant attributes against inflammation, free radicals, and apoptosis. This study investigated the likely protective property of hesperetin against cisplatin-induced testicular and epididymal toxicity. Forty mice randomly assigned into four groups of 10 animals each were administered with the appropriate regimen. Group 1 (control) received normal saline alone while group 2 received 2.5 mg/kg body weight of cisplatin only. Group 3 received 40 mg/kg body weight of hesperetin and 2.5 mg/kg body weight of cisplatin, while group 4 received only 40 mg/kg body weight of hesperetin. Hesperetin administration (orally) was for 30 consecutive days while cisplatin (intraperitoneally) was on days 5, 7, 10, 13, 17, 20, 24, and 28. Findings revealed that cisplatin decreased the activities/level of glutathione peroxidase, superoxide dismutase, reduced glutathione, nitric oxide, total thiol, testosterone, luteinizing hormone, total cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol, sperm viability, sperm motility, and total sperm count while the abnormal sperm cells and levels of lipid peroxidation, H2O2, and inteleukin-1β were increased (P < 0.05). Furthermore, histopathological results of examined testes showed a reduction in the tube-shaped size of testes, diminished development of spermatozoa, and architectural impairment with cisplatin exposure. Contrarily, hesperetin reversed cisplatin-induced testicular and epididymal oxidative damage, suppressed steroidogenesis and spermatogenesis, and repaired the architecture of the testes. The study showed that hesperetin demonstrated multi-mechanistic protective activity against testicular dysfunction, oxidative stress, and histological damage owing to its antioxidant and steroidogenic actions. [ABSTRACT FROM AUTHOR]

Introdction: Human spermatogenesis is a complex process that transforms spermatogonial stem cells through mitosis and meiosis to spermatozoa. Testosterone is the key regulator of the terminal stages of meiosis, adherence of spermatids to Sertoli cells, and spermiation. Follicle‐stimulating hormone (FSH) may be required for early spermatogenesis and is important for maintaining normal spermatogenesis in men. Hormonal contraception suppresses FSH, luteinizing hormone, and intratesticular testosterone concentration, resulting in marked suppression of sperm output. Results: Clinical trials using testosterone alone or testosterone plus progestin demonstrate that sustained suppression of sperm concentration to ≤1 million/mL is sufficient to prevent pregnancy in the female partner. New agents that target spermatogenesis could use this as a target for contraceptive efficacy while others that block sperm function or transport may require a lower threshold. When sperm concentrations are suppressed to such low levels, measurement of sperm motility and morphology is technically difficult and unnecessary. With current data from fertile and infertile men, it is not possible to establish a lower limit of sperm motility or percent normal morphology that equates to the prevention of conception. New compounds that decrease sperm motility or alter sperm morphology may need to demonstrate a complete absence of sperm motility or altered morphology in all spermatozoa in the ejaculate. Sperm function tests may be useful depending on the mechanism of action of each new compound. Conclusion: Monitoring of sperm surrogate markers to ensure effective contraception relies on laboratories experienced in semen analyses. The development of at‐home tests to assess sperm parameters has progressed rapidly. Some tests have been assessed in clinical trials and approved by regulatory agencies for at‐home use for fertility assessment. However, caution must be exercised in using these tests as many have not been rigorously validated against semen parameters measured in laboratories by trained technologists using standardized tests defined in the World Health Organization Semen Manual. [ABSTRACT FROM AUTHOR]

Purpose: Infertility is a global health issue and nutrition plays a significant role in fertility outcomes. We aimed to investigate the cross-sectional and prospective associations of glycemic index (GI) and glycemic load (GL) with semen quality parameters in a cohort of healthy young men. Materials and Methods: The study included 106 men aged 18–35 years from the FERTINUTS trial. Dietary intake was estimated through 3-day dietary records and several semen parameters were assessed. Multivariable linear regression analysis with the Least Absolute Shrinkage and Selection Operator (LASSO) approach was employed. Results: The cross-sectional analysis revealed positive associations between GI and GL and total sperm count, sperm concentration, and total motility. In the prospective analysis, baseline GI was associated with increases in pH, vitality, immotile sperm or abnormal midpiece and decreases in total sperm count and motility. Conversely, GL was positively associated with changes in vitality and total sperm count. Conclusions: While these findings suggest that GI may have adverse effects on several sperm quality parameters, the results were not consistently observed in the cross-sectional analysis. However, GL was consistently associated with better sperm quality in both analyses. The impact of carbohydrate quality and quantity on fertility remains uncertain and larger prospective studies are needed. [ABSTRACT FROM AUTHOR]

Hyaluronan (HA) is a glycosaminoglycan that forms a gel-like barrier in the subcutaneous (SC) space, limiting bulk fluid flow and the dispersion of SC-administered therapeutics. Recombinant human hyaluronidase PH20 (rHuPH20) facilitates the rapid delivery of co-administered therapeutics by depolymerizing HA in the SC space. Administration of rHuPH20 can induce the formation of anti-rHuPH20 antibodies, or anti-drug antibodies (ADAs), with the potential to bind endogenous PH20 hyaluronidase in the adult testes and epididymis. Using a variety of relevant animal models and multiple dose regimens of rHuPH20 across the full spectrum of animal development, we demonstrated that rHuPH20 administration resulted in the formation of ADAs. Although these ADAs can bind both the recombinant rHuPH20 enzyme and recombinant versions of animal model-specific hyaluronidases, they had no impact on fertility parameters (as measured by sperm concentration and motility, litter size, and litter viability) or fetal development. We present the result of our nonclinical studies in order of the developmental lifecycle, beginning with adults. Toxicology studies that extend beyond the standard package are also presented. These studies demonstrate the favorable safety profile of rHuPH20 and ADAs in nonclinical models. Additionally, we identified substantial safety margins for clinically relevant doses of rHuPH20. [Display omitted] • rHuPH20 facilitates subcutaneous administration of therapeutics. • Impact of rHuPH20 and anti-drug antibodies observed in multiple nonclinical models. • rHuPH20 anti-drug antibodies did not negatively alter fertility or development. • Substantial safety margins were identified when compared to human-equivalent doses. • Studies support favorable safety profile of rHuPH20. [ABSTRACT FROM AUTHOR]

Male infertility due to asthenozoospermia is quite frequent, but its etiology is poorly understood. We recruited two infertile brothers, born to first‐cousin parents from Pakistan, displaying idiopathic asthenozoospermia with mild stuttering disorder but no ciliary‐related symptoms. Whole‐exome sequencing identified a splicing variant (c.916+1G>A) in ARMC3, recessively co‐segregating with asthenozoospermia in the family. The ARMC3 protein is evolutionarily highly conserved and is mostly expressed in the brain and testicular tissue of human. The ARMC3 splicing mutation leads to the exclusion of exon 8, resulting in a predicted truncated protein (p.Glu245_Asp305delfs*16). Quantitative real‐time PCR revealed a significant decrease at mRNA level for ARMC3 and Western blot analysis did not detect ARMC3 protein in the patient's sperm. Individuals homozygous for the ARMC3 splicing variant displayed reduced sperm motility with frequent morphological abnormalities of sperm flagella. Transmission electron microscopy of the affected individual IV: 2 revealed vacuolation in sperm mitochondria at the midpiece and disrupted flagellar ultrastructure in the principal and end piece. Altogether, our results indicate that this novel homozygous ARMC3 splicing mutation destabilizes sperm flagella and leads to asthenozoospermia in our patients, providing a novel marker for genetic counseling and diagnosis of male infertility. [ABSTRACT FROM AUTHOR]

Little is known about the effects of low gravity on human gametes. The aim of this study was to analyze if fresh human sperm samples after fast gravity load changes suffered any detrimental effect in comparison to the splits maintained in Earth's gravity. Fifteen fresh samples from normozoospermic donors were analyzed. Statistically significant differences in vitality (69.7 ± 9.9 % vs 72.4 ± 9.7 %, [95 % CI: 0.002,0.07]); motile sperm concentration (23.7 ± 15.3 M/ml vs 31.5 ± 25.1 M/ml, [95 % CI: 1.03,14.65]); grade "a" sperm concentration (8.7 ± 6.5 M/ml vs 11.7 ± 9.9 M/ml, [95 % CI: 0.71,5.28]); percentage of progressive motility sperm (30 ± 12.9 % vs 36 ± 14.3 %, [95 % CI: 0.10,0.37]) and curvilinear velocity VCL: 45.7 ± 12.8 μm/s vs 47.7 ± 13.3 μm/s, [95 % CI: 0.79,3.22]) were observed. No statistical differences were observed in other sperm kinematic parameters, morphology, DNA fragmentation, apoptosis, and oxidative stress. In conclusion, even though it did not result in a total loss, heavy gravity load changes including microgravity causes a significant decrease in sperm vitality and motility suggesting that negative consequences would be even higher if the exposure were longer. The results obtained indicate that further research is really needed before Assisted Reproduction will be considered for the future human reproduction outside the Earth. • Motility and Vitality of human sperm are significantly decreased after parabolic flight including short periods of microgravity. • Curvilinear Velocity of human male gametes was particularly decreased. • No changes were found in other sperm kinematic parameters, morphology, DNA fragmentation, apoptosis and oxidative stress. • Results were found after short exposure; suggesting more negative consequences regarding human fertility in a longer time exposure. • More studies needed before Assisted Reproduction Techniques can be considered for the likelihood of Human Reproduction in Space. [ABSTRACT FROM AUTHOR]

The decrease in fertility in aging roosters is related to the reduced quality of ejaculated sperm. This study aimed to investigate the effect of dietary supplementation with ginseng extract at various concentrations (0–150 mg/kg) on testicular function, semen preservation, and fertility at different stages of sexual maturity (mature and aging roosters) in Thai native roosters. Pradu Hang Dum roosters at 32 (mature; n = 24) and 75 (aging; n = 24) weeks of age were fed diets with non-supplemented or supplemented ginseng extracts (50, 100, and 150 mg/kg) until the end of the experiment. In experiment 1, fresh semen samples were examined for the quality parameters of semen volume, sperm concentration, sperm motility, sperm viability, lipid peroxidation, and enzymatic activities. In experiment 2, semen was preserved at 5 °C for up to 48 h, and the semen quality and fertility potential were determined. In experiment 3, testicular function and testosterone concentrations were evaluated. The results showed that ginseng extract supplementation in the diets of both mature and aging roosters at 50 and 100 mg/kg improved fresh semen quality (P < 0.05). A decrease in malondialdehyde levels in fresh semen was observed with increasing enzyme activities. In mature roosters, the progressive motility of cold-stored semen and fertility rates were higher in the G50 and G100 groups compared to the control and G150 groups after 24 h of storage (P < 0.05). In aging roosters, the highest significant differences in progressive motility, viability, and fertility rates were observed in the G50 and G100 groups at all storage times (P < 0.01). These improvements might be attributed to good testicular function in spermatogenesis, as revealed by the results of histological examination and testosterone concentrations. However, higher doses of ginseng extract supplementation negatively affected sperm quality. In summary, the recommended dose of ginseng extract supplementation in diets is 50 mg/kg. Fertility results indicated that insemination with semen preserved for 24 h was satisfactory in both mature and aging roosters. • Ginseng extract improved sperm production and semen preservation in mature and aging roosters. • Semen quality was highest in G50 and G100 groups, confirmed by histological and testosterone results. • High doses (150 mg/kg) negatively impacted sperm quality. • 24 hour preserved semen showed satisfactory fertility rates in both mature and aging roosters. [ABSTRACT FROM AUTHOR]

Parabens (PBs) are widely used in the cosmetic, pharmaceutical, and food industries as preservatives of products. Because of its great use, humans and other organisms are highly exposed daily. However, little is known about the effect of PBs on male infertility. Therefore, the aim of the present study was to evaluate the effect of methylparaben (MePB) and propylparaben (PrPB), alone or in combination, on the physiological characteristics of pig in vitro exposed sperm to different concentrations (0, 200, 500, and 700 μM) for viability, motility, and acrosome integrity evaluation and (0, 200, 500, 700, 1000, and 2000 μM) for DNA fragmentation index evaluation, after 4 h of exposure. The results showed that sperm viability decreased after exposure to MePB from the concentration of 500 μM. In the PrPB and mixture groups, viability decreased at all concentrations except for the control. The decrease in viability of sperm exposed to PrPB was greater than that of the mixture and MePB groups. Sperm motility decreased in all the experimental groups exposed to PBs, at all concentrations, except for the control group. Acrosome integrity was not decreased in the MePB group; however, in the PrPB group, it decreased at a concentration of 200 μM and in the mixture at 500 μM. All groups exhibited DNA damage at different concentrations, except for the control group. Additionally, the effect of PBs on sperm quality was concentration‐dependent. The results demonstrated that MePB and PrPB alone or in combination can have adverse effects on sperm quality parameters. MePB had lower toxicity than did both PrPB and the mixture. The mixture did not have an additive effect on any of the parameters evaluated. This could partially explain the link between PB exposure and infertility. Sperm viability decreased after exposure to MePB from the concentration of 500 μM. In the PrPB and mixture groups, viability decreased at all concentrations. Sperm motility decreased in all the experimental groups exposed to PBs at all concentrations. Acrosome integrity was not decreased in the MePB group; however, in the PrPB group, it decreased at a concentration of 200 μM and in the mixture at 500 μM. All groups exhibited DNA damage at different concentrations. [ABSTRACT FROM AUTHOR]

Background: Valproic acid (VPA) is a widely used antiepileptic drug (AED) often prescribed as a first-line treatment for many idiopathic and symptomatic generalized epilepsies. Several studies have highlighted the side effects of VPA on male fertility and reproductive factors in males, although the specific underlying etiology of these abnormalities is not clear. The present systematic review and meta-analysis aimed to assess the preclinical and clinical evidence concerning the impact of VPA on male fertility and reproductive factors. Methods: The scientific literature was reviewed for eligibility using PubMed, Web of Science, and PsycINFO, encompassing preclinical and clinical studies. Factors related to male fertility and reproduction, such as differences in sperm count, sperm motility, and the percentage of abnormal sperm, were compared between the experimental groups treated with VPA (in both preclinical and clinical) and the control groups using the Standardized Mean Difference (SMD) with 95% confidence intervals (CIs). Additionally, differences in follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were explicitly assessed in clinical studies. Results: Male fertility data were extracted from 7 preclinical studies (112 animals) and 5 clinical studies (274 male individuals). The results of animal studies found that the sperm count (SMD = -2.28, 95% CI: -3.39 to -1.18, P = 0.335) and sperm motility (SMD = -2.32, 95% CI: -3.34 to -1.30, P = 0.368) were decreased in the treated groups compared to the control groups. The percentage of abnormal sperm (SMD = 3.27, 95% CI: 1.98 to 4.56, P = 0.019) was significantly increased, while a non-significant reduction was revealed in the weight of the testis (SMD = -2.73, 95% CI: -4.23 to -1.23, P = 0.673) in treated groups. The outcomes of clinical studies indicated a non-significant decrease in sperm count (SMD = -0.78, 95% CI: -1.58 to 0.03, P = 0.286) among patients with epilepsy treated with VPA compared to control subjects. However, a significant reduction in sperm motility (SMD = -1.62, 95% CI: -2.81 to -0.43, P = 0.033 was observed. The percentage of abnormal sperm showed a non-significant increase (SMD = 0.93, 95% CI: -0.97 to 2.84, P = 0.616) after being treated with VPA. Furthermore, there was a non-significant reduction in the levels of FSH (SMD = -1.32, 95% CI: -2.93 to 0.29, P = 0.198) and LH (SMD = -0.96, 95% CI: -1.95 to 0.04, P = 0.211) observed in clinical participants. Conclusion: This meta-analysis of both preclinical and clinical studies revealed that VPA causes a significant reduction in male fertility and reproductive factors among male patients with epilepsy. Clinical neurologists should be more cautious when prescribing VPA, especially to young male adult patients with epilepsy. [ABSTRACT FROM AUTHOR]

Background Objective Materials and Methods Results Discussion and Conclusions Primary spermatogenic disorders represent a severe form of male infertility whereby sperm production is impaired due to testicular dysfunction, leading to reduced quality or quantity of spermatozoa. Gene‐centered research has certainly demonstrated the importance of the genetic factor in the etiology of both poor sperm morphology or motility and reduced sperm count. In the last decade, next‐generation sequencing has expanded the research to whole exome which has transformed our understanding of male infertility genetics, but uncertainty persists in its diagnostic yield, especially in large unrelated populations.To evaluate the utility of exome sequencing in detecting genetic factors contributing to various traits of primary spermatogenic disorders, which is a crucial step before interpreting the diagnostic yield of the platform.We manually curated 415 manuscripts and included 19 research studies that predominantly performed whole exome sequencing in cohorts of unrelated cases with primary spermatogenic defects.The detection rate, defined as the fraction of cases with an identifiable genetic cause, typically remained below 25% for quantitative defects of spermatozoa, whereas improved rates were observed for traits of abnormal sperm morphology/motility and in populations enriched with consanguineous families. Unlike the quantitative defects, the genetic architecture of the qualitative issues of spermatozoa featured a small number of recurrent genes describing a large fraction of studied cases. These observations were also in line with the lower biological complexity of the pathways affected by the reported genes.This review demonstrates the variability in detection rates of exome sequencing across semen phenotypes, which may have an impact on the expectations of the diagnostic yield in the clinical setting. [ABSTRACT FROM AUTHOR]

Simple Summary: Liquid-preserved boar semen is widely used in the pig breeding industry. Preservation and transportation can affect the fertilizing ability of sperm, reducing the efficiency of artificial insemination. Betaine, a natural plant extract, has been shown to benefit male fertility. However, it remains unclear whether it exerts a protective effect on liquid boar sperm during preservation and transportation. The results showed that the addition of appropriate amounts of betaine to boar semen extenders effectively mitigated the adverse effects of road transport and room temperature storage on sperm quality. Specifically, betaine supplementation showed significant protective effects against temperature fluctuation and vibration stress that inevitably occurred during transportation. The increased antioxidant capacity of boar sperm may underlie its beneficial effects. These findings offer a theoretical foundation for the use of betaine in artificial insemination procedures. Boar semen is commonly used in artificial insemination (AI) for pig breeding, but its quality can be negatively affected by liquid preservation and transportation, leading to reduced fertility rates. Vibration and temperature fluctuations are critical factors that significantly impact semen quality during storage and transportation, influencing the success rate of AI procedures. Betaine, a naturally occurring compound known for its role in maintaining male fertility, demonstrates potential for improving the preservation and transportation of liquid-preserved boar sperm. The present study demonstrated that betaine supplementation in the semen extenders at 0.5 mg/mL had a significant protective effect on boar sperm motility during storage at 17 °C for 3 to 5 days. During road transportation, 2.5 mg/mL betaine showed significant protective effects on boar sperm progressive motility, while 0.4 mg/mL betaine notably improved boar sperm mitochondrial activity and antioxidant capacity, and reduced lipid peroxidation damage. Simulation models also demonstrated that betaine supplementation increased the proportion of sperm displaying progressive motility and possessing intact acrosomes, regardless of the storage temperature (17 °C or 25 °C), and effectively mitigated the damage caused by vibration at a speed of 200 r/min. Overall, supplementing liquid-preserved boar semen extenders with betaine shows promise in mitigating damage to sperm quality during storage and transportation. [ABSTRACT FROM AUTHOR]

Simple Summary: Spermatozoa cryopreservation technology has been widely used to preserve high-quality male spermatozoa and has to some extent solved the problem of spermatozoa supply in yellow catfish (Pelteobagrus fulvidraco). However, cryopreservation can still cause cellular damage and affect spermatozoa viability and fertility. Therefore, it is necessary to study cryo-injury, which is beneficial for further optimizing the spermatozoa cryopreservation protocol. In this experiment, we investigated the effects of adding or not adding 10% DMSO on spermatozoa motility, ultrastructural damage, oxidative damage, and DNA damage of thawed yellow catfish. Using 10% DMSO as a cryoprotectant can improve spermatozoa motility (75.64 ± 1.12), significantly alleviate injuries to organelles and membrane structures, improve SOD (7.99 × 104 ± 5.24 nmol/mgprot) and T-AOC (3.7 × 105 ± 0.06 nmol/mgprot) enzyme activity, reduce MDA (1.09 ± 0.45 nmol/mgprot) enzyme activity, and mitigate the degree of oxidative damage. It also greatly reduces the DNA fragmentation rate (59 ± 1.56%). The research results provide data support for the optimization of the cryopreservation protocol for the spermatozoa of yellow catfish, which is beneficial for the preservation of the germplasm resources of yellow catfish and the effective utilization of resources during large-scale production. Spermatozoa cryopreservation protocols have been established for yellow catfish (Pelteobagrus fulvidraco), but cryopreservation can still cause cellular damage and affect spermatozoa viability and fertility. Therefore, the aim of this paper was to evaluate the effects of adding or not adding cryoprotectants during low-temperature storage on the ultrastructural damage, oxidative damage, and DNA damage of thawed yellow catfish spermatozoa. The mixed semen of three male yellow catfish was divided into a fresh spermatozoa group, a frozen spermatozoa group (DMSO+) with a cryoprotectant (10% DMSO), and a frozen spermatozoa group without a cryoprotectant (DMSO−). Ultrastructural of the spermatozoa after thawing were observed under an electron microscope and the spermatozoa were assayed for SOD, MDA, and T-AOC enzyme activities, as well as for DNA integrity. In terms of movement parameters, compared with DMSO−, the addition of DMSO has significantly improved sperm motility, curve line velocity (VCL), and straight line velocity (VSL). The ultrastructural results showed that although thawed spermatozoa exhibited increased damage than fresh spermatozoa, 10% DMSO effectively reduced the damage to the plasma membrane, mitochondria, and flagellum of spermatozoa by cryopreservation, and most of the spermatozoa were preserved with intact structure. The results of oxidative damage showed that compared with frozen spermatozoa, 10% DMSO significantly increased the activities of SOD and T-AOC enzymes and clearly reduced the activity of the MDA enzyme. The antioxidant capacity of spermatozoa was improved, lipid peroxidation was reduced, and the oxidative damage caused by cryopreservation was mitigated. The DNA integrity of spermatozoa showed that 10% DMSO clearly reduced the DNA fragmentation rate. In conclusion, 10% DMSO can effectively reduce the ultrastructural damage, oxidative damage, and DNA damage of yellow catfish spermatozoa during cryopreservation; it can also further optimize the cryopreservation protocol for yellow catfish spermatozoa. Meanwhile, it also provides a theoretical basis for the future optimization of the cryopreservation protocols. [ABSTRACT FROM AUTHOR]

Mature, vital, and motile spermatozoa are essential for reaching the oocyte and binding to hyaluronic acid (HA) in the cumulus oophorus matrix. This study aims to determine the relationship between sperm-migration ability and HA-binding potential, as well as the relationship between sperm concentration and motility. Semen samples were collected from 702 men aged 20–56 years (median 34.8). We evaluated the sperm concentration and motility from basic semen analysis, the swim-up test (expressed as millions per mL and the migration efficiency percentage), and the hyaluronan-binding assay (HBA). A moderate positive correlation was found between the migration test results and HBA (R = 0.48). The highest correlation was observed between the concentration of motile spermatozoa and the migration test results (R = 0.85) and HBA (R = 0.4). The sperm migration efficiency strongly correlated with progressive motility (R = 0.6). Although significantly higher sperm migration was observed in patients with normal HBA results, the results of the functional tests were found to differ in some cases. For infertility treatment, the current diagnostic algorithm should be enhanced with more comprehensive seminological methods that assess the sperm-migration ability and HA-binding potential. We also recommend incorporating the swim-up method into the diagnostic protocol before planning assisted reproductive technology (ART) treatment. [ABSTRACT FROM AUTHOR]

Introduction Materials and Methods Results Conclusions Microsurgical testicular sperm extraction (microTESE) is crucial for treating non‐obstructive azoospermia (NOA), offering both ‘fresh’ and ‘frozen’ options. This study evaluates the impact of fresh versus frozen microTESE on the progression to intra‐cytoplasmic sperm injection (ICSI) cycles, focusing on sperm motility.We conducted a retrospective analysis of microTESE procedures at a major medical centre from 2007 to 2021, excluding cases of obstructive azoospermia and cryptozoospermia. Patients were divided into two groups: fresh microTESE (Group FR) and frozen microTESE (Group FZ). Sperm motility was assessed, and ICSI outcomes were compared between groups.Out of 128 microTESE procedures on 113 NOA patients, 31 were fresh and 97 were frozen. Sperm was found in 67.7% of fresh cases and 45.3% of frozen cases. In fresh cases, 85.7% had motile sperm for ICSI, whereas in frozen cases, 81.8% had motile sperm initially, but only 52.7% retained motility post‐thaw.Our findings indicate a significant drop in motile sperm availability for ICSI in frozen microTESE cases compared to fresh ones. This suggests a potential advantage of fresh microTESE for certain couples, despite the logistical challenges, highlighting the need for careful patient selection and counselling. [ABSTRACT FROM AUTHOR]

Context: Prostate-specific antigen (PSA) is important for semen liquefaction and sperm motility. Anti-PSA antibodies may lead to immune infertility. Aims: The present study was conducted to evaluate the impact of seminal anti-PSA antibodies on semen parameters in fertile and infertile men. Methods: A cross-sectional analytic study was conducted on 105 fertile men (≥21–50 years) having biological children (within last 2 years) with normal semen analysis as controls and 105 infertile men with abnormal semen analysis as cases. All semen samples were cryopreserved till 210 samples were collected, followed by estimation of anti-PSA antibodies using an indirect enzyme-linked immunosorbent assay technique. Key results: Mean ± standard deviation (s.d.) age of 210 participants was 30.0 ± 4.65 years. Mean ± s.d. levels of seminal anti-PSA antibodies in infertile men were 27.82 ± 102.19 ng/mL and in fertile men −30.45 ± 49.49 ng/mL (P = 0.001). A significant negative correlation was observed between anti-PSA antibody levels and sperm concentration (P = 0.013), rapid progressive motility (P = 0.001), slow progressive motility (P = 0.006), progressive sperm motility (P = 0.001), and normal morphology (P = 0.001), and significant positive correlation was observed with immotile sperms (P = 0.001). The overall accuracy of anti-PSA antibody for differentiating infertile from fertile men was 63.33%. Conclusions: Seminal anti-PSA antibodies were significantly correlated with semen parameters in fertile and infertile men with an accuracy of 63.33%. A negative correlation was observed between antibody levels and progressive sperm motility. Implications: Seminal anti-PSA antibodies can be used as a biomarker for male infertility assessment. Globally, male factor infertility is increasing and so is the need for newer and effective tests for its diagnosis and management. The present study was conducted to evaluate the impact of seminal anti-prostate-specific antibodies (PSA) antibodies on semen parameters in fertile and infertile men. A significant negative correlation was observed with sperm concentration, motility, and morphology, and a positive correlation was observed with grade D motility. The overall accuracy of anti-PSA antibodies in predicting infertile from fertile men was 63.33%. Image by Naina Kumar. [ABSTRACT FROM AUTHOR]

Melatonin, a hormone synthesized in various tissues, plays a crucial role in modulating sperm characteristics, yet its protective function on boar sperm remains poorly understood. This study aimed to investigate the expression and localization of melatonin-related proteins (AANAT, ASMT, MT1, MT2, and NQO2) in pig tissues, assess the impact of melatonin on pig sperm motility parameters and quality, and elucidate the underlying molecular mechanisms. Our results revealed widespread expression of AANAT, ASMT, MT1, MT2, and NQO2 proteins in pig tissues, particularly in the testis. Specific localization patterns were observed in Leydig cells, reproductive epithelium, and columnar epithelium cells in the testis and cauda epididymis. Additionally, melatonin membrane receptors MT1 and MT2 were detected in boar sperm. Melatonin treatment significantly enhanced boar sperm motility parameters and quality, particularly with 10 nM melatonin treatment. Inhibition of the MT1 receptor, but not the MT2 receptor, resulted in decreased sperm motility, highlighting the pivotal role of the MT1 receptor in mediating melatonin's effects on boar sperm. Metabolomic analysis revealed significant alterations in sperm metabolites following melatonin supplementation, particularly in amino acid metabolism. Overall, our findings provide comprehensive insights into melatonin's mechanisms in improving boar sperm quality, suggesting its potential as a therapeutic agent for enhancing male fertility. • Comprehensive study shows melatonin boosts boar sperm motility and quality, mainly via the MT1 receptor. • Melatonin-related proteins in pig tissues suggest roles in testicular physiology and sperm protection. • Melatonin alters seminal plasma metabolites, showing diverse impacts on sperm function and metabolism. [ABSTRACT FROM AUTHOR]

Background: Centrifugation is a common procedure to improve the quality of chilled and frozen canine semen by removing debris and seminal plasma and adding semen extenders. The aim of this study was to evaluate the efficacy and influence of a second centrifugation after 48 h of storage at 5 °C on the sperm quality of canine semen. The ejaculates of 45 healthy male dogs, divided into three groups according to body weight, were analyzed for macro- and microparameters such as ejaculate volume, sperm concentration, kinematic parameters, morphology, and integrity of plasma membrane. Samples were analyzed at baseline conditions (T0), after 24 h (T24) and after 48 h (T48) to assess the effects of the different treatments on sperm quality. Results: The results showed a significant effect of a second centrifugation on the improvement of chilled sperm quality compared to the other techniques, especially up to 48 h. Conclusions: Analysis of the data showed that the semen samples centrifuged and then cooled at 5 °C had acceptable semen parameters, especially in terms of motility, with a gradual decrease in serial evaluations after 24 and 48 h. A second centrifugation after 48 h of storage may lead to better semen quality and improve the kinetics of sperm parameters, the percentage of morphologically normal sperm and the percentage of sperm with intact membranes. [ABSTRACT FROM AUTHOR]

Myo‐inositol may be efficient to improve sperm parameters to increase the chance of fertility. Although, the data are controversial. This study aimed to assess the impact of Myo‐inositol supplements on semen quality and in vitro fertilization (IVF) outcomes. In this systematic review and meta‐analysis, a comprehensive search was conducted in PubMed, Web of Science, and Embase. The objective was to identify relevant human studies that investigated the effects of Myo‐inositol treatment on various sperm factors, such as sperm motility, sperm concentration, sperm morphology, viable spermatozoa, spermatozoa with DNA fragmentation, and pregnancy rate. Additionally, the testosterone levels of patients with Oligo‐astheno‐teratozoospermia (OAT) after Myo‐inositol application were considered. The findings of 16 selected studies from 2240 citations indicated significant improvements in several parameters of sperm after Myo‐inositol administration. Myo‐inositol treatment was associated with a notable increase in total sperm motility (SMD 0.90; 95% CI: 0.34 to 1.46; I2 = 0%, p = .001) and progressive sperm motility (SMD 1.48; 95% CI: 0.37 to 2.59; I2  = 0%, p = .008). Additionally, there was a significant improvement in testosterone levels (SMD 0.54; 95% CI: 0.34 to 0.73; I2  = 0%, p < .0001). Furthermore, Myo‐inositol therapy demonstrated a significant decrease in spermatozoa with DNA fragmentation (SMD −1.37; 95% CI: −2.43 to −0.32; I2  = 85%, p = .01). This study suggests that Myo‐inositol therapy has a positive impact on specific sperm parameters, such as total and progressive sperm motility, along with testosterone levels. These findings provide support for the potential benefits of Myo‐inositol in improving male fertility parameters related to sperm factors. [ABSTRACT FROM AUTHOR]

Semen quality and fertility has declined over the last 50 years, corresponding to ever-increasing environmental stressors. However, the cellular mechanisms involved and their impact on sperm functions remain unknown. In a repeated sampling human cohort study, we identify a significant effect of prior perceived stress to increase sperm motility 2-3 months following stress, timing that expands upon our previous studies revealing significant stress-associated changes in sperm RNA important for fertility. We mechanistically examine this post-stress timing in mice using an in vitro stress model in the epididymal epithelial cells responsible for sperm maturation and find 7282 differentially H3K27me3 bound DNA regions involving genes critical for mitochondrial and metabolic pathways. Further, prior stress exposure significantly changes the composition and size of epithelial cell-secreted extracellular vesicles that when incubated with mouse sperm, increase mitochondrial respiration and sperm motility, adding to our prior work showing impacts on embryo development. Together, these studies identify a time-dependent, translational signaling pathway that communicates stress experience to sperm, ultimately affecting reproductive functions. Changes in semen quality over the last 50 years correspond with increasing environmental stressors. Here, Moon et al. causally identified stress-mediated changes in secreted extracellular vesicles as regulators of sperm motility in mice and men. [ABSTRACT FROM AUTHOR]

Background Objective Materials and methods Results Conclusion Recently, the metabolic pathways involved in energy production and the role of aquaglyceroporins in capacitation‐associated events have been studied in humans and mice. However, little is known about these in ram spermatozoa.The present study investigated bioenergetic and aquaglyceroporin 3 variations during in vitro capacitation of ram spermatozoa. In addition, differences in testosterone levels between males were examined to determine their influence on capacitation‐like changes.Spermatozoa obtained from nine rams (ejaculates = 36) were incubated for 180 min in three different media (control, capacitating, and aquaglyceroporin‐inhibitor media) at 38.5°C. At 0 and 180 min of incubation in each medium, sperm viability, kinetics, chlortetracycline patterns, adenosine triphosphate concentration, lactate excretion (final subproduct of glycolysis), and immunolocalization of aquaporin 3 were evaluated.The increment of the capacitated spermatozoa‐chlortetracycline pattern and the hyperactivated‐like movement characterized by the highest curvilinear velocity and amplitude of lateral head displacement and the lowest linearity was only recorded after 180 min in the capacitating medium. At this time and conditions, adenosine triphosphate content and lactate excretion decreased, whereas the aquaglyceroporin 3 location in the midpiece and principal piece increased compared to 0 min. Such changes were not observed in the control medium over time. Incubation in the aquaglyceroporin‐inhibitor medium for 180 min reduced drastically sperm motility and adenosine triphosphate content compared to the other media. Testosterone analysis revealed a significant individual variability, which was also present in all sperm parameters evaluated. Furthermore, testosterone was negatively correlated with adenosine triphosphate content but positively correlated with lactate excretion levels, sperm viability, motility, capacitated sperm‐chlortetracycline pattern, and aquaglyceroporin 3 immunolabeling in the midpiece and principal piece.Despite individual differences, capacitation of ram spermatozoa increases adenosine triphosphate consumption, energy metabolism, and aquaglyceroporin 3 location in the midpiece and principal piece, which seems to be related to the acquisition of hyperactivated‐like motility. Furthermore, testosterone levels may serve as a valuable tool to select those males with a greater sperm metabolism rate and fertilizing capacity. [ABSTRACT FROM AUTHOR]

Introduction: Over the past few decades, there has been growing concern about the concurrent trends of increasing Autism Spectrum Disorder (ASD) prevalence and declining sperm quality. These trends represent significant public health challenges that warrant thorough investigation of their underlying causes and implications. Objectives: The primary objectives of this study are to analyze trends in ASD prevalence and sperm quality parameters from 2000 to 2024, assess the statistical significance and effect size of these trends, explore potential correlations between ASD prevalence and sperm quality parameters, and identify significant predictors among sperm quality parameters that influence ASD prevalence. Methods: This study employed a longitudinal approach using multiple regression, time series analysis, ANOVA, Principal Component Analysis (PCA), hierarchical clustering, logistic regression, and cross-correlation analysis. Data on ASD prevalence were sourced from the CDC Autism and Developmental Disabilities Monitoring Network, while sperm quality data were collected from various published studies. Results: The findings reveal significant negative associations between ASD prevalence and sperm quality parameters such as sperm concentration and motility, suggesting that better sperm quality is linked to lower ASD rates. Conversely, parameters like sperm DNA fragmentation (SDF), volume of ejaculate, pH level, and semen viscosity show positive associations with ASD prevalence, indicating higher values in these parameters correlate with higher ASD rates. Conclusion: The study highlights the importance of maintaining reproductive health to potentially mitigate ASD risk and calls for further research to elucidate the underlying mechanisms driving these trends. These findings support the hypothesis that reproductive health factors play a crucial role in ASD etiology and suggest potential biological markers for assessing ASD risk. [ABSTRACT FROM AUTHOR]

Heat stress is a serious problem that affects animal husbandry by reducing growth and reproductive performance of animals. Adding plant extracts to the diet is an effective way to help overcome this problem. Alginate oligosaccharide (AOS) is a natural non-toxic antioxidant with multiple biological activities. This study analyzed the potential mechanism of AOS in alleviating heat stress and improving semen quality in boars through a combination of multiple omics tools. The results indicated that AOS could significantly increase sperm motility (P < 0.001) and sperm concentration (P < 0.05). At the same time, AOS improved the antioxidant capacity of blood and semen, and increased blood testosterone (P < 0.05) level. AOS could improve the metabolites in sperm, change the composition of gut microbiota, increase the relative abundance of beneficial bacteria such as Pseudomonas (P < 0.01), Escherichia-Shigella (P < 0.05), Bifidobacterium (P < 0.01), reduce the relative abundance of harmful bacteria such as Prevotella_9 (P < 0.05), Prevotellaceae_UCG-001 (P < 0.01), and increase the content of short chain fatty acids. Proteomic results showed that AOS increased proteins related to spermatogenesis, while decreasing heat shock protein 70 (P < 0.05) and heat shock protein 90 (P < 0.01). These results were verified using immunofluorescence staining technology. There was a good correlation among sperm quality, sperm metabolome, sperm proteome, and gut microbiota. In conclusion, AOS can be used as a feed additive to increase the semen quality of boars to enhance reproductive performance under heat stress. [ABSTRACT FROM AUTHOR]

Despite sharing an autosomal genome, the often divergent reproductive strategies of males and females cause the selection to act in a sex-specific manner. Selection acting on one sex can have negative, positive, or neutral fitness consequences on the opposite sex. Here, we test how female-limited selection on reproductive investment in Japanese quail (Coturnix japonica) affects male fertility-related traits. Despite there being no difference in the size of males' testes from lines selected for high female reproductive investment (H-line) or low female reproductive investment (L-line), in both lines, the left testis had a greater volume of sperm-producing tissue. Since H-line females have a larger left-side restricted oviduct, this suggests a positive genetic correlation between male and female gonad function and that internal testis structure is a target of sexual selection. However, despite H-line males having previously been found to have greater fertilization success in a competitive scenario, we found little evidence of a difference between the lines in sperm number, motility, velocity, length, or the number of sperm that reached the ova. Precopulatory cues and/or the role of seminal fluid in sperm motility may thus be more likely to contribute to the H-line male fertilization advantage in this species. Graphical Abstract [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effectiveness of hesperidin (HES) on testicular histopathological changes, biochemical changes, and semen characteristics in rats exposed to glyphosate (GLP). The control group was given a normal diet devoid of GLP and HES, the HES group was given 100 mg/kg/day HES with the normal diet, the GLP group was given GLP at the LD50/10 dose of normal feed, which was 787.85 mg/kg/day, and the GLP + HES group was given normal feed containing 787.85 mg/kg/day LD50/10 dose of GLP in addition to 100 mg/kg/day HES. GLP administration reduced sperm motility, sperm plasma membrane integrity, glutathione levels, and total antioxidant levels in the testicular tissues of rats. Moreover, it caused an increase in right testis and left epididymis weights, abnormal sperm counts, malondialdehyde levels, total oxidant status, and DNA damage. The HES treatment showed curative effects on these parameters. Furthermore, HES was effective in lessening the histopathological damage that was caused by GLP. The results showedthat HES protects spermatological parameters and DNA integrity, improves antioxidant defenses, and lowers the damage and lipid peroxidation caused by GLP in testicular tissue. [ABSTRACT FROM AUTHOR]

Simple Summary: When a couple faces infertility challenges like sperm motility issues, they often visit fertility clinics for assistance. Poor sperm motility can delay fertilization, making natural conception difficult. In an andrology laboratory, embryologists test the semen sample. Before assisted reproduction procedures, embryologists usually perform a group of steps to prepare the semen sample and to isolate the best sperms. These technical steps might harm the sperms, because they become susceptible to something so called; oxidative stress, which occurs when too many harmful molecules called free radicals accumulate in the test tube, leading to cell damage and death. A group of chemicals called antioxidants can neutralize these free radicals. The researchers in this study investigated the therapeutic effects of Mitoquinone, which is a mitochondria-targeted antioxidant, on the motility of human sperms exposed to oxidative stress during in vitro procedures. Mitoquinone as an antioxidant might have therapeutic potential for sperms' oxidative-damage-associated disorders. This investigation paves the route for future research to confirm the beneficial role of Mitoquinone supplementation in improving and enhancing sperm quality during in vitro processes in the andrology laboratories. The preparation of human sperm in an andrology laboratory subjects it to oxidative stress. Reactive oxygen species are produced by mitochondria, making it susceptible to oxidative damage; hence, mitochondria-targeted antioxidants like Mitoquinone (MitoQ) might have therapeutic potential for oxidative-damage-associated disorders. The current research aims to establish whether MitoQ has any positive effects during in vitro preparation of fresh human sperm. Viability and motility are evaluated to determine the effective MitoQ concentration and to assess whether MitoQ supplementation is affected by sperm concentration by incubating normospermia semen samples at 37 °C for 2 h and 4 h, respectively. The effect of semen centrifugation following supplementation of 20 × 106 sperm/mL with 200 nM MitoQ is also assessed by measuring viability, motility and sperm DNA fragmentation. The best sperm motility is achieved after 2 h of incubation with 200 nM MitoQ at 37 °C. Sperm concentration of 20 × 106 sperm/mL is the best concentration where 200 nM MitoQ works efficiently. For semen centrifugation at 300× g for 20 min, supplementation with 200 nM MitoQ shows higher sperm motility. The current results demonstrate that MitoQ supplementation during in vitro human semen preparation procedures positively affects fresh sperm motility without affecting viability or increasing DNA fragmentation. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the effect of a coconut water-based extender on the quality parameters of dog semen (n=24 ejaculates) during refrigeration (4°C) preservation up to 72 h. Four ejaculates per dog were collected from 6 dogs by digital manipulation. All fresh ejaculates were evaluated for macroscopic and microscopic semen evaluation tests. The semen samples were then extended in a coconut water-based extender using a split sample technique. The diluted semen was kept in a refrigerator for preservation at 4°C and evaluated at 0, 24, 48, and 72 h for sperm quality parameters. The sperm motility significantly reduced over a period of time from 0 to 72 h. The live sperm percentage did not differ significantly at 0, 24 and 48 h of preservation. However, there was a significant drop in live sperm percentage at 72 h. The percent sperm abnormalities were significantly increased from 24, 48 to 72 h of preservation. In coconut water-based extender there was a significant difference in intact plasma membrane percentage at different time slots (0, 24, 48 and 72 h). It was concluded that coconut water-based extender can preserve canine sperm for up to 48 h while maintaining sperm motility and viability above 50%. Sperm abnormalities remained within 20% and plasma membrane integrity above 60% both were maintained up to 72 h of semen preservation. [ABSTRACT FROM AUTHOR]

The addition of antioxidants to cryopreservation media reportedly improves sperm post‐thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L‐carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post‐thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA–glucose–based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk–based extender with 20% egg yolk and 5% dimethylformamide (milk control); (3) Extender 1 supplemented with 50 mM L‐carnitine and 10 mM pyruvate (EDTA‐carnitine‐pyruvate); and (4) Extender 2 supplemented with 50 mM L‐carnitine and 10 mM pyruvate (milk‐carnitine‐pyruvate). In Experiment 2, 50 mM L‐carnitine and 10 mM pyruvate were added post‐thaw to samples cryopreserved with extenders 1 and 2 (EDTA control and milk control). Sperm kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability were evaluated after thawing. No significant differences (p > 0.05) were observed for most of the kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability of spermatozoa, between the samples frozen in the presence or absence of L‐carnitine and pyruvate, nor between the samples after the post‐thaw addition of these components. A higher (p < 0.05) mean velocity and higher (p < 0.05) amplitude of lateral head displacement were observed in the samples frozen in the milk‐based extender with the addition of L‐carnitine and pyruvate after thawing. The addition of 50 mM L‐carnitine and 10 mM pyruvate, either to the freezing extenders or after thawing, was not deleterious for sperm; however, it did not improve equine sperm motility, viability, acrosome and DNA integrity, nor decrease membrane lipid peroxidation after thawing. [ABSTRACT FROM AUTHOR]

Fertility control has traditionally been applied in zoos to control captive populations, and reversible contraception is important. However, contraceptive methods for male bears have not been reported. We aimed to establish a reversible contraceptive for male brown bears by investigating the effects of a gonadotropin‐releasing hormone (GnRH) vaccine (Improvac®) that was developed for the immune castration of pigs. We vaccinated six bears with two sequential doses of 400 (n = 2) or 600 μg Improvac® (n = 4) with a 1‐month interval during the pre‐breeding season (February to April). We compared the reproductive parameters (testosterone levels and semen parameters) of the six vaccinated and four non‐vaccinated (control) bears once during the breeding season (May or June). To investigate whether the reproductive performance could be restored in the following year of contraception, we also compared the reproductive parameters once during the breeding season in two bears between the year with GnRH vaccination and the following year without vaccination. Vaccination treatments suppressed reproductive parameters in 5 bears, although vaccination with 400 μg of Improvac® was not effective in one bear. Testosterone levels and the rate of progressive sperm motility were significantly lower, and total sperm count and testis size tended to be lower in vaccinated bears, compared with the controls. Blood biochemical findings and direct observations after Improvac® vaccination did not reveal side effects. Moreover, testosterone levels and spermatogenic scores of two bears were restored in the following year. We confirmed that the Improvac® vaccine elicited a reversible contraceptive effect in male brown bears. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the results of semen analysis using the manual method and the SQA-Vision sperm analyser after four years of practice and with a large cohort of patients. This was a comparative study of 1130 cases collected for semen analysis between October 2019 and October 2023, which were analysed simultaneously and independently by different operators using the manual microscopic method and an SQA-V automated analyser. For each sample, sperm concentration, progressive motility, motility, normal morphology, and round cells count were performed. There was no significant difference between the SQA-V method and manual assessment for all sperm parameters (Mann–Whitney test p > 0.05). According to the parameter studied, there was a strong correlation (rho = 0.81) and a very high correlation (rho = 0.98) between manual assessment and the SQA-V method. In the analysis of sperm concentration, the sensitivity and specificity were 0.90 and 0.99, respectively. The sensitivity and specificity for the analysis of sperm progressive motility were 0.98 and 0.99, respectively, while the sensitivity and specificity for the analysis of sperm motility were 0.87 and 0.99, respectively. The sensitivity and specificity for the analysis of normal morphology were 0.88 and 0.99, respectively. Regarding the analysis of round cells, the sensitivity and specificity were 0.98 and 0.99, respectively. The results of this retrospective study indicate that the SQA-V system offers satisfactory performance for routine sperm analysis. [ABSTRACT FROM AUTHOR]

Proper connection between the sperm head and tail is critical for sperm motility and fertilization. Head–tail linkage is mediated by the head-tail coupling apparatus (HTCA), which secures the axoneme (tail) to the nucleus (head). However, the molecular architecture of the HTCA is poorly understood. Here, we use Drosophila to investigate formation and remodeling of the HTCA throughout spermiogenesis by visualizing key components of this complex. Using structured illumination microscopy, we demonstrate that key HTCA proteins Spag4 and Yuri form a 'centriole cap' that surrounds the centriole (or basal body) as it invaginates into the surface of the nucleus. As development progresses, the centriole is laterally displaced to the side of the nucleus while the HTCA expands under the nucleus, forming what we term the 'nuclear shelf'. We next show that the proximal centriole-like (PCL) structure is positioned under the nuclear shelf, functioning as a crucial stabilizer of centriole-nucleus attachment. Together, our data indicate that the HTCA is a complex, multi-point attachment site that simultaneously engages the PCL, the centriole and the nucleus to ensure proper head-tail connection during late-stage spermiogenesis. [ABSTRACT FROM AUTHOR]

Purpose: To identify novel variants in ACTL9 and new phenotypes responsible for male infertility. Methods: Genomic DNA was extracted from peripheral blood samples for whole-exome sequencing (WES). Computer-assisted sperm analysis (CASA) was used to test the motility of spermatozoa. The ultrastructure of flagella and the mitochondrial sheath were assessed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Immunostaining was used to validate the localization and expression of ACTL9 and ACTL7A. An Actl9-mutated mouse model was used to validate the phenotypes by CASA and TEM. Results: We identified novel homozygous variants in ACTL9 in two independent Chinese families. Spermatozoa with ACTL9 mutations showed decreased CASA parameters and a higher proportion of spermatozoa with abnormal morphology, exhibiting coiled flagella and a thickened midpiece. The spermatozoa were characterized by chaotic or irregular '9+2' structures and irregular mitochondrial sheath arrangements in the flagellum. Actl9 knock-in mice also showed abnormal CASA parameters and irregular '9+2' structures in flagella. Conclusions: Our study expands the mutation spectrum and phenotypic spectrum of ACTL9. [ABSTRACT FROM AUTHOR]

Spermatozoon volume regulation is an essential determinant of male fertility competence in mammals and oviparous fishes. In mammals, aquaporin water channels (AQP3, -7 and -8) have been suggested to play a role in spermatozoon cell volume regulatory responses in the hypotonic female oviduct. In contrast, the ejaculated spermatozoa of marine teleosts, such as the gilthead seabream (Sparus aurata), experience a high hypertonic shock in seawater, initially resulting in an Aqp1aa-mediated water efflux, cell shrinkage and the activation of motility. Further regulatory recovery of cell volume in post-activated spermatozoa is mediated by Aqp4a in cooperation with the Trpv4 Ca2+ channel and other ion channels and transporters. Using a paralog-specific antibody, here, we show that seabream spermatozoa also express the aquaglyceroporin AQP3 ortholog Aqp3a, which is highly accumulated in the mid posterior region of the spermatozoon flagella, in a similar pattern to that described in mouse and human sperm. To investigate the role of Aqp3a in seabream sperm motility, we used a recently developed AQP3 antagonist (DFP00173), as well as the seabream Aqp3a-specific antibody (α-SaAqp3a), both of which specifically inhibit Aqp3a-mediated water conductance when the channel was heterologously expressed in Xenopus laevis oocytes. Inhibition with either DFP00173 or α-SaAqp3a did not affect sperm motility activation but did impair the spermatozoon motion kinetics at 30 s post activation in a dose-dependent manner. Interestingly, in close resemblance to the phenotypes of AQP3-deficient murine sperm, electron microscopy image analysis revealed that both Aqp3a inhibitors induce abnormal sperm tail morphologies, including swelling and angulation of the tail, with complete coiling of the flagella in some cases. These findings suggest a conserved role of Aqp3a as an osmosensor that regulates cell volume in fish spermatozoa under a high hypertonic stress, thereby controlling the efflux of water and/or solutes in the post-activated spermatozoon. [ABSTRACT FROM AUTHOR]

Simple Summary: This study compares two methods of preparing boar sperm for in vitro fertilization: the traditional density gradient selection (DGS) using centrifugation and the newer microfluidic chip-based sperm (MCS) sorting. MCS resulted in lower sperm concentration and fewer morphologically abnormal sperm compared to DGS. Additionally, although DGS showed higher progressive motility, both methods had similar fertilization outcomes in terms of cleavage rates, blastulation rates, and embryo quality. The results show that MCS performs at least as well as DGS and may offer advantages due to its gentler approach and potential for more consistent results in IVF procedures. Porcine in vitro embryo production (IVP) protocols have conventionally used density gradient selection (DGS) by centrifugation to prepare sperm samples and achieve successful fertilisation. However, the possible toxicity of the solutions used and the potential damage caused by the centrifugation step may have a negative effect on the quality of the sample. Microfluidic chip-based sperm (MCS) sorting has been proposed as an alternative technique for the selection of high-quality sperm with the purpose of improving reproductive outcomes in IVF. This device does not require centrifugation or any toxic solution to prepare the sample for fertilisation. The sample is not subjected to unnecessary stress, and the process is less operator-dependent. In this study, we compared the sperm parameters of unselected extender-diluted boar semen samples with selected samples using DGS and MCS methods. The results show an expected reduction in sperm concentration after both methods. All the groups were significantly different from one another, with MCS being the group with the lowest concentration. Though the three groups had a similar overall motility, significant differences were found in progressive motility when comparing the unselected group (control, 19.5 ± 1.4%) with DGS and MCS. Progressive motility in DGS was also significantly higher than in MCS (65.2 ± 4.9% and 45.7% ± 5.3, respectively). However, MCS selection resulted in enriched sperm samples with a significantly lower proportion of morphologically abnormal sperm compared to DGS. After fertilisation, no statistical differences were found between the two methods for embryological parameters such as cleavage rates, blastulation rates, and embryo quality. The number of cells in blastocysts derived from MCS was significantly greater than those derived from DGS sperm. Thus, we demonstrate that MCS is at least as good as the standard DGS for most measures. As a more gentle and reproducible approach for sperm selection, however, it could improve consistency and improve IVP outcomes as mediated by a greater proportion of morphologically normal sperm and manifested by a higher cell count in blastocysts. [ABSTRACT FROM AUTHOR]

Background: Sperm cryopreservation potential can be explored in diverse conditions to hamper future fertility. Reactive oxygen species (ROS) were among the prime candidate responsible for vulnerability of the sperms due to lack of intracellular antioxidants defense. Aim: The study aim to investigate the association between DNA fragmentation index and antioxidant enzyme. Methods: This controlled observational hospital based study was conducted on men participants (n=100). The semen specimens with normal sperm count and motility post fertility work up was collected in a sterile vial containing cryoprotectant and thereafter stored in a freezer having liquid nitrogen -196 °C for 7 days at controlled rate. Sperm morphology, DNA fragmentation and motility along with malondialdehyde and total antioxidant capacity (TAC) were measured before and after cryopreservation. Results: Semen specimens post cryopreservation demonstrated significant decrease in sperm motility, number and malondialdehyde level (p<0.001). There was a significant increase in DNA fragmentation and total antioxidant capacity levels (p<0.001). Conclusion(s): Sperm cryopreservation may lead to deleterious variations of structure and function(s) due to generation of ROS that contribute in DNA fragmentation. [ABSTRACT FROM AUTHOR]

The study is the first to formulate and investigate potential of papaya seed chloroform extract based solid lipid nanoparticles (PSCEN) as antifertility agents on male Bandicota bengalensis. The prepared nanoparticles were spherical of size 300–600 nm. The release kinetics showed a controlled release of the drug with major release over 48 h. To assess the antifertility effects of PSCEN, adult male rats were fed a diet containing two different concentrations of PSCEN (5% and 10%) for 15 days under bi-choice conditions. The mean total active ingredient ingestion of the rats in the two treated groups ranged from 2.13–3.31 and 3.92–5.87 g/100g body weight, respectively. No adverse effects of treatment on body weight were observed. Also, no mortality of rats was observed. The treatment had a significant effect on the weight of the testis and the epididymis, but not on the other organs. Sperm motility (%), sperm viability (%), sperm count (millions/ml), sperm mitochondrial activity (%), sperm nuclear chromatin de-condensation (%) and sperm hypo-osmotic swelling (%) were significantly decreased, and sperm abnormality (%) significantly increased compared to the vehicle control group. The reproductive success rates of male rats treated with 5% and 10% PSCEN and mated with untreated female rats were 20.00–66.67% and 16.67%, respectively, while in untreated female rats mated with male rats of vehicle control group, reproductive success rate was 33.33 to 80%. The study found a maximal antifertility effect of the 10% PSCEN containing bait, which was irreversible up to 105 days after stopping treatment, suggesting long-term efficacy. [ABSTRACT FROM AUTHOR]

The effects of honey addition to Tris media on osmolality, pH values and on the quality characteristics of fresh and chilled goat spermatozoa were studied. Semen samples were collected from five Shami bucks and incubated at 20 °C for the fresh samples and at 5 °C for the chilled ones in Tris-Based Medium (TBM) and Tris-Egg Yolk (TEY) medium without (controls) and with honey at ratios of 1%, 2% and 4% (v/v) respectively. Osmolality and pH levels were determined for all used media. Spermatozoa motility was analysed using the Computer Assisted Sperm Analyzer (CASA) system and the integrity of sperm plasma membrane was tested by the Hypo-Osmotic Swelling Test (HOST). Osmolality values were significantly increased (P<0.05) by honey addition reaching 558 mOsm/kg in TBM and 675 mOsm/kg in TEY with 4% of honey compared with 307 mOsm/kg and 312 mOsm/kg for the controls respectively. In contrast, pH values decreased (P<0.05) from 6.93 in TBM control medium to 6.4 in TBM with 4% of honey and from 6.83 in TEY control medium to 6.1 in TEY with 4% of honey. CASA motility parameters including the percent motility (MOT%) and the percent of sperm showing progressive motility (PMOT%) increased after adding 1% and 2% of honey while the addition of 4% led to a clear decrease (P<0.05) for the fresh and chilled samples. The percentages of spermatozoa subpopulations including the rapid % and the static % categories for fresh and chilled samples were significantly decreased and increased respectively after adding 4% honey. Moreover, for all media and for all incubation temperatures, the HOST percentage decreased significantly (P<0.05) by adding 4% honey. It was concluded that honey addition to Tris media influenced the osmolality and pH values, which in turn affected the quality characteristics of buck spermatozoa based on honey concentration regardless of the incubation temperature or the used media. [ABSTRACT FROM AUTHOR]

With a prospect to improve goat artificial insemination, this study aims to evaluate the effects of storage time, egg yolk concentration and semen washing on the rate of motile spermatozoa (spz) in Arbia breed bucks semen stored at 5 °C for 48 h. Semen was collected from three mature bucks using an artificial vagina during December 2018. The ejaculates used in this study (rate of motile sperm > 75 %) were divided into two fractions: A, and B, (unwashed and washed samples, respectively): themselves divided into three fractions, and diluted in Tris-Acid citric based extenders, containing egg yolk at 2 %, 10 % and 20 %. ANOVA test was performed using the R software version 3.3.0. The rate of motile spermatozoa was evaluated after 24 and 48 hours of storage in the refrigerator at 5 °C. It registered a significant decrease as a function of the storage time in the different extenders. It varied between 42.43 ± 9.86 % and 61.29 ± 5.44 %, after 24 hours of storage, and between 35.86 ± 13.95 %, and 55.45 ± 11.65 % after 48 hours of storage. The semen washing significantly reduced the rate of motile spermatozoa in the different extenders. Increasing the concentration of egg yolk in the extender increased the rate of motile spermatozoa, especially during the first 24 hours of storage. In conclusion, the rate of spermatozoa motility of unwashed semen stored at 5 °C is higher in extenders containing high egg-yolk concentration, and decrease according to conservation time. [ABSTRACT FROM AUTHOR]